There are two objectives to the research. One objective is to decipher how a promoter functions. The approach is to manipulate a promoter sequence and alter the nucleotide bases in a carefully controlled pattern so as to change the ability of control proteins and RNA polymerase to interact with this promoter. The method for completing this objective involves chemical and enzymatic synthesis of the promoter and, most importantly, sequence variants of the promoter. Specifically this objective will focus on either the total lac promoter or the major rightward control region of bacteriophage lambda. A choice remains to be made. Depending upon which system is selected, we want to probe in precise biochemical and chemical terms how CAP functions to attenuate transcription, how the directionality and initiation site for E. coli RNA polymerase transcription is defined, and how cro protein, E. coli RNA polymerase, and cI repressor interact cooperatively and antagonistically on the same DNA. Additional studies will concentrate on how individual control proteins recognize and interact with specific promoter sequences. These combined studies will contribute significantly toward an understanding of gene regulation at the molecular level. The second objective is to develop rapid methods for the synthesis of deoxyoligonucleotides. The strength of our approach for studying gene regulation is dependent on manipulating the DNA sequence and studying the effect of these changes on transcriptional control. We do not want our research to be limited by the rate of chemical synthesis and therefore have a need to improve this methodology.